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transport of coenzyme m (2-mercaptoethanesulfonic acid) in methanobacterium ruminantium.a system for transport of coenzyme m, 2-mercaptoethanesulfonic acid (hs--com), in methanobacterium ruminatium strain m1 required energy, showed saturation kinetics, and concentrated the coenzyme against a gradient. the process was sensitive to temperature and was maximally active at ph 7.1. cells took up hs--com at a linear rate, with a vmax of 312 pmol/min per mg (dry weight) and an apparent km of 73 nm. an intracellular pool of up to 5 mm accumulated which was not exchangeable with the medium. ...197933148
distribution of coenzyme f420 and properties of its hydrolytic fragments.the ability of hydrolytic products of coenzyme f420 to substitute for f420 in the hydrogenase and nicotinamide adenine dinucleotide phosphate-liniked hydrogenase systems of methanobacterium strain m.o.h. was kinetically determined. the nicotinamide adenine dinucleotide phosphate-linked hydrogenase system was employed to quantitate the levels of f420 in a number of methanogenic bacteria as well as in some nonmethanogens. methanobacterium ruminantium and methanosarcina barkeri contained low levels ...197940952
specificity and biological distribution of coenzyme m (2-mercaptoethanesulfonic acid).the specificity of the growth requirement of methanobacterium ruminantium strain m1 for a new coenzyme, 2-mercaptoethanesulfonic acid (hs--com), was examined. a variety of derivatives, analogs, and potential biosynthetic precursors of coenzyme m were tested; only a restricted range of thioether, thioester, and thiocarbonate derivatives of the cofactor were found to replace the hs--com requirement. bromoethanesulfonic acid (brch2ch2so3-), a halogenated analog of hs--com, potently inhibited the gr ...1979104960
an ancient divergence among the bacteria.the 16s ribosomal rnas from two species of methanogenic bacteria, the mesophile methanobacterium ruminantium and the thermophile methanobacterium thermoautotrophicum, have been characterized in terms of the oligonucleotides produced by digestion with t1 ribonuclease. these two organisms are found to be sufficiently related that they can be considered members of the same genus or family. however, they bear only slight resemblance to "typical" procaryotic genera; such as escherichia, bacillus and ...1977408502
influence of ch4 production by methanobacterium ruminantium on the fermentation of glucose and lactate by selenomonas ruminantium.a method is described for increasing the production of h2 from glucose or lactate by selenomonas ruminantium by sequential transfers in media containing pregrown methanobacterium ruminantium. the methanogen uses the h2 formed by the selenomonad to reduce co2 to ch4. analysis of fermentation products from glucose showed that lactate was the major product formed from glucose by s. ruminantium alone. several sequential transfers in the presence of the methanogen caused a marked decrease in lactate ...1977596874
presence of growth factors for methanobacterium ruminantium in both gram-positive and gram-negative bacteria. 19744219266
coenzyme m, essential for growth of a rumen strain of methanobacterium ruminantium.a growth factor present in rumen fluid and essential for growth of a rumen strain of methanobacterium ruminantium was shown to be coenzyme m, 2-mercaptoethanesulfonic acid.19744376145
characteristics of s organism isolated from methanobacillus omelianskii.previous work showed that methanobacillus omelianskii was a mixed culture of an ethanol-oxidizing organism called s organism and a hydrogen-utilizing methane bacterium, strain moh. s organism grows poorly on ethanol unless a hydrogen-utilizing methanogenic bacterium is included to utilize the h(2) produced during growth. further studies have shown that, among many substrates tested, only ethanol, n-propanol, n-butanol, isobutanol, n-pentanol, acetaldehyde, oxalacetate, and pyruvate are fermented ...19725058442
formate as an intermediate in the bovine rumen fermentation.an average of 11 (range, 2 to 47) mumoles of formate per g per hr was produced and used in whole bovine rumen contents incubated in vitro, as calculated from the product of the specific turnover rate constant, k, times the concentration of intercellular formate. the latter varied between 5 and 26 (average, 12) nmoles/g. the concentration of formate in the total rumen contents was as much as 1,000 times greater, presumably owing to formate within the microbial cells. the concentration of formate ...19705419259
isolation and characterization of methanobacterium ruminantium n. sp. 195813549377
phylogenetic analysis of methyl coenzyme-m reductase detected from the bovine rumen.the object of the present study is isolation of methyl coenzyme-m reductase (mcr) genes (mcra) from the bovine rumen fluid and determination of phylogenetical placements of the genes to investigate mechanisms of methanogenesis in the rumen from a point of view of mcra genes.200415287871
sediment distribution of methanogenic bacteria in lake erie and cleveland harbor.the direct fluorescent-antibody technique was employed to determine the distribution patterns of four species of methanogens in the sediments of lake erie and cleveland harbor. methanobacterium ruminantium was the most numerous methanogen found in regions of high-organic-silt sediments. the population of this species ranged from 10 to 10 cells/g of dry sediment. methanobacterium strain moh and methanosarcina barkeri were identified in sand-silt, clay, or sand sediments. these methanogens ranged ...198016345529
anaerobic transformation of furfural by methanococcus deltae (delta)lh.methanococcus deltae (delta)lh was grown on h(inf2)-co(inf2) in the presence of various concentrations of furfural. furfural at higher concentrations, namely, 20 and 25 mm, inhibited growth of this organism. at concentration of 5 and 10 mm, no inhibition of growth was observed. the other methanogens in this study were not inhibited by 10 mm furfural. among the methanogens tested, m. deltae was capable of transforming furfural, whereas methanobacterium thermoautotrophicum marburg, methanosarcina ...199716535618
postinoculation protozoan establishment and association patterns of methanogenic archaea in the ovine rumen.association patterns between archaea and rumen protozoa were evaluated by analyzing archaeal 16s rrna gene clone libraries from ovine rumen inoculated with different protozoa. five protozoan inoculation treatments, fauna free (negative control), holotrich and cellulolytic protozoa, isotricha and dasytricha spp., entodinium spp., and total fauna (type a) were tested. we used denaturing gradient gel electrophoresis, quantitative pcr, and phylogenetic analysis to evaluate the impact of the protozoa ...200717513586
molecular diversity of methanogens in feedlot cattle from ontario and prince edward island, canada.the molecular diversity of rumen methanogens in feedlot cattle and the composition of the methanogen populations in these animals from two geographic locations were investigated using 16s rrna gene libraries prepared from pooled pcr products from 10 animals in ontario (127 clones) and 10 animals from prince edward island (114 clones). a total of 241 clones were examined, with methanobrevibacter ruminantium accounting for more than one-third (85 clones) of the clones identified. from these 241 cl ...200717483285
analysis of methanogen diversity in the rumen using temporal temperature gradient gel electrophoresis: identification of uncultured methanogens.a temporal temperature gradient gel electrophoresis (ttge) method was developed to determine the diversity of methanogen populations in the rumen. tests with amplicons from genomic dna from 12 cultured methanogens showed single bands for all strains, with only two showing apparently comigrating bands. fingerprints of methanogen populations were analyzed from dna extracted from rumen contents from two cattle and four sheep grazing pasture. for one sheep, dilution cultures selective for methanogen ...200717431710
application of pseudomurein endoisopeptidase to fluorescence in situ hybridization of methanogens within the family methanobacteriaceae.in situ detection of methanogens within the family methanobacteriaceae is sometimes known to be unsuccessful due to the difficulty in permeability of oligonucleotide probes. pseudomurein endoisopeptidase (pei), a lytic enzyme that specifically acts on their cell walls, was applied prior to 16s rrna-targeting fluorescence in situ hybridization (fish). for this purpose, pure cultured methanogens within this family, methanobacterium bryantii, methanobrevibacter ruminantium, methanosphaera stadtmana ...200616950902
anaerobic degradation of lactate by syntrophic associations of methanosarcina barkeri and desulfovibrio species and effect of h(2) on acetate degradation.when grown in the absence of added sulfate, cocultures of desulfovibrio desulfuricans or desulfovibrio vulgaris with methanobrevibacter smithii (methanobacterium ruminantium), which uses h(2) and co(2) for methanogenesis, degraded lactate, with the production of acetate and ch(4). when d. desulfuricans or d. vulgaris was grown in the absence of added sulfate in coculture with methanosarcina barkeri (type strain), which uses both h(2)-co(2) and acetate for methanogenesis, lactate was stoichiometr ...198116345708
effect of monensin and lasalocid-sodium on the growth of methanogenic and rumen saccharolytic bacteria.it is thought that monensin increases the efficiency of feed utilization by cattle by altering the rumen fermentation. we studied the effect of monensin and the related ionophore antibiotic lasalocid-sodium (hoffman-laroche) on the growth of methanogenic and rumen saccharolytic bacteria in a complex medium containing rumen fluid. ruminococcus albus, ruminococcus flavefaciens, and butyrivibrio fibrisolvens were inhibited by 2.5 mug of monensin or lasalocid per ml. growth of bacteroides succinogen ...197916345418
effects of several inhibitors on pure cultures of ruminal methanogens.to examine the effects of five inhibitors of methanogenesis, 2-bromoethanesulphonate (bes), 3-bromopropanesulphonate (bps), lumazine, propynoic acid and ethyl 2-butynoate, on ch4 production of the ruminal methanogens methanobrevibacter ruminantium, methanosarcina mazei and methanomicrobium mobile.200415281932
phylogenetic analysis of methanogens from the bovine rumen.interest in methanogens from ruminants has resulted from the role of methane in global warming and from the fact that cattle typically lose 6 % of ingested energy as methane. several species of methanogens have been isolated from ruminants. however they are difficult to culture, few have been consistently found in high numbers, and it is likely that major species of rumen methanogens are yet to be identified.200111384509
effects of the fusarium spp. mycotoxins fusaric acid and deoxynivalenol on the growth of ruminococcus albus and methanobrevibacter ruminantium.the fusarium spp. mycotoxins fusaric acid and deoxynivalenol (don) were tested for antimicrobial activity against ruminococcus albus and methanobrevibacter ruminantium. the growth of both organisms was inhibited by fusaric acid as low as 15 micrograms/ml (84 microm) but not by don, at levels as high as 100 micrograms/ml (338 microm). no synergistic inhibitory effect was observed with don plus fusaric acid. neither organism was able to adapt to the fusaric acid and responses of each organism to t ...200010941514
phylogenetic study of methanogens associated with rumen ciliates.the phylogeny of methanogenic archaea associated with ciliate protozoa in a sheep rumen was investigated. ruminal ciliate protozoa were exhaustively washed and mixtures of genomic dna extracted. archaea-specific nested pcr amplification was conducted with the ciliate genomic mixture. the resultant small subunit (16s) ribosomal rna gene (ssu rdna) was cloned into escherichia coli jm 109. many methanogens were still observed on and/or in ciliate cells by fluorescent microscopy even after exhaustiv ...199910441724
syntrophic association of a butyrate-degrading bacterium and methanosarcina enriched from bovine rumen fluid.an anaerobic butyrate-degrading bacterium, morphologically similar to syntrophomonas wolfei, was isolated in coculture with desulfovibrio strain g11 from an enrichment of bovine rumen fluid. a methanosarcina species was the major h2-using organism in the enrichment. the results are discussed in relationship to the absence of methanospirillum hungatei, the h2-using methanogen usually found in association with s. wolfei, and the finding of methanosarcina rather than methanobrevibacter ruminantium ...19817224635
rapidly growing rumen methanogenic organism that synthesizes coenzyme m and has a high affinity for formate.methanogenic bacteria with a coccobacillus morphology similar to methanobrevibacter ruminantium were isolated from the bovine rumen. one isolate, 10-16b, represented a previously undescribed rumen population that, unlike m. ruminantium, synthesized coenzyme m, grew rapidly (mu = 0.24 h-1) on h2-co2 in a complex medium, had simple nutritional requirements, and metabolized formate at reported rumen concentrations. h2 was metabolized to partial pressures 10-fold lower than those reported for the ru ...19846433795
hydrogen-oxidizing methane bacteria. ii. electron microscopy.comparison of the fine structure of the methanogenic organism from the culture known as methanobacillus omelianskii with that of methanobacterium formicicum revealed a great similarity. both organisms exhibited a large number of intracytoplasmic membranous elements when stained with phosphotungstic acid. in contrast, no such elements were observed in methanobacterium ruminantium.19685643051
characteristics of methanogens isolated from bovine rumen.six strains of methanogens were isolated from 10(-8) and 10(-9) ml of bovine rumen contents. all strains had the morphologic and physiologic characteristics of methanobrevibacter spp. four strains required coenzyme m; two did not. growth of all strains either depended on or was stimulated by a mixture of isobutyric, isovaleric, 2-methylbutyric, and valeric acids. none of the strains reacted with antiserum against the type strain of methanobrevibacter ruminantium.19863954338
is coenzyme m bound to factor f430 in methanogenic bacteria? experiments with methanobrevibacter ruminantium.coenzyme m (2-mercaptoethane sulfonic acid) and factor f430 (a nickel porphinoid) are coenzymes found in methanogenic bacteria. recently it has been proposed that in these bacteria a coenzyme mf430 also exists which plays a key role in methane formation and in which coenzyme m and f430 are bound to each other. to test this hypothesis methanobrevibacter ruminantium, which requires coenzyme m as a vitamin, was grown in the presence of [2-14c]comsh. f430 and 'com' (mixture of comsh and its disulfid ...19853920049
new approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (hs-com)-dependent growth of methanobacterium ruminantium in a pressureized atmosphere.the sensitivity of the requirement of methanobacterium ruminantium strain m1 to a new coenzyme, 2-mercaptoethanesulfonic acid (hs-com) was examined by use of new techniques that were developed for rapid and efficient handling of large numbers of cultures of methanogenic bacteria. the system uses sealed tubes that contain a gas mixture of 80% hydrogen and 20% carbon dioxide under a pressure of 2 to 3 atm. this modification of the hungate technique reduces variability among replicate cultures and ...1976827241
inhibitory effects of h2 on growth of clostridium cellobioparum.hydrogen inhibits the growth of hydrogen-producing clostridium cellobioparum, but not of escherichia coli or bacteroides ruminicola. the inhibition is reversible. when hydrogen was removed either by palladium black or by gassing out the tube, glucose utilization increased as did optical density and hydrogen production of c. cellobioparum. removal of the h2 by methanogenic bacteria favors the growth of c. cellobioparum. grown with methanobacterium ruminantium in various concentrations of glucose, ...1976779644
methane formation and methane oxidation by methanogenic bacteria.methanogenic bacteria were found to form and oxidize methane at the same time. as compared to the quantity of methane formed, the amount of methane simultaneously oxidized varied between 0.3 and 0.001%, depending on the strain used. all the nine tested strains of methane producers (methanobacterium ruminantium, methanobacterium strain m.o.h., m. formicicum, m. thermoautotrophicum, m. arbophilicum, methanobacterium strain az, methanosarcina barkeri, methanospirillum hungatii, and the "acetate org ...1979762019
fermentation of cellulose by ruminococcus flavefaciens in the presence and absence of methanobacterium ruminantium.the anaerobic cellulolytic rumen bacterium ruminococcus flavefaciens normally produces succinic acid as a major fermentation product together with acetic and formic acids, h2, and co2. when grown on cellulose and in the presence of the methanogenic rumen bacterium methanobacterium ruminantium, acetate was the major fermentation product; succinate was formed in small amounts; little formate was detected; h2 did not accumulate; and large amounts of ch4 were formed. m. ruminantium depends for growt ...1977562131
methane synthesis without the addition of adenosine triphosphate by cell membranes isolated from methanobacterium ruminantium.the membrane fraction isolated from broken cells of methanobacterium ruminantium actively synthesized methane from co2 and h2 without the addition of atp or other cofactors. this activity was lost unless strictly anaerobic conditions were maintained throughout the isolation and incubation procedures. 3h2, but not 3h2o, was readily incorporated into methane. this indicates that hydrogen atoms are used in the formation of methane without the prior equilibration of protons with the water phase. met ...1979435275
factor 420-dependent pyridine nucleotide-linked formate metabolism of methanobacterium ruminantium.methanobacterium ruminantium was shown to possess a formate dehydrogenase which is linked to factor 420 (f420) as the first low-molecular-weight or anionic electron transfer coenzyme. reduced f420 obtained from the formate dehydrogenase can be further linked to the formation of hydrogen via the previously described f420-dependent hydrogenase reaction, thus constituting an apparently simple formate hydrogenlyase system, or to the reduction of nicotinamide adenine dinucleotide phosphate via f420:n ...1975234935
factor 420-dependent pyridine nucleotide-linked hydrogenase system of methanobacterium ruminantium.methanobacterium ruminantium was shown to possess a nicotinamide adenine dinucleotide phosphate (nadp)-linked factor 420 (f420)-dependent hydrogenase system. this system was also shown to be present in methanobacterium strain moh. the hydrogenase system of m. ruminantium also links directly to f420, flavin adenine dinucleotide (fad), flavin mononucleotide (fmn), methyl viologen, and fe-3 plus. it has a ph optimum of about 8 and an apparent km for f420 of about 5 x 10-6 m at ph 8 when nadp is the ...1975234934
rumen-like methanogens identified from the crop of the folivorous south american bird, the hoatzin (opisthocomus hoazin).the hoatzin is the only known avian species with foregut fermentation. it is a primarily folivorous feeder and has a distended crop and lower/distal esophagus, which has evolved for the microbial fermentation of ingested feed. crop samples collected from 10 individual animals from the apure river area, apure state, venezuela were examined for the presence and density of methanogens using 16s rrna gene clone libraries and real-time pcr prepared from pooled and individual pcr products. a total of ...200919387486
effect of tea saponin on methanogenesis, microbial community structure and expression of mcra gene, in cultures of rumen micro-organisms.to determine the in-vitro effect and mode of action of tea saponin on the rumen microbial community and methane production.200819146532
methanobrevibacter ruminantium as an indicator of domesticated-ruminant fecal pollution in surface waters.a pcr-based assay (mrnif) targeting the nifh gene of methanobrevibacter ruminantium was developed to detect fecal pollution from domesticated ruminants in environmental water samples. the assay produced the expected amplification product only when the reaction mixture contained dna extracted from m. ruminantium culture, bovine (80%), sheep (100%), and goat (75%) feces, and water samples from a bovine waste lagoon (100%) and a creek contaminated with bovine lagoon waste (100%). the assay appears ...200717827308
the genome sequence of the rumen methanogen methanobrevibacter ruminantium reveals new possibilities for controlling ruminant methane emissions.methane (ch(4)) is a potent greenhouse gas (ghg), having a global warming potential 21 times that of carbon dioxide (co(2)). methane emissions from agriculture represent around 40% of the emissions produced by human-related activities, the single largest source being enteric fermentation, mainly in ruminant livestock. technologies to reduce these emissions are lacking. ruminant methane is formed by the action of methanogenic archaea typified by methanobrevibacter ruminantium, which is present in ...201020126622
development of a vaccine to mitigate greenhouse gas emissions in agriculture: vaccination of sheep with methanogen fractions induces antibodies that block methane production in vitro.to develop an understanding of the immune responses of ruminants to methanogens, and to provide proof of a concept that harnessing the immune system of ruminants is a potentially viable approach to mitigate greenhouse gas emissions from agriculture.201020200573
methanobrevibacter millerae sp. nov. and methanobrevibacter olleyae sp. nov., methanogens from the ovine and bovine rumen that can utilize formate for growth.four formate-utilizing methanogens were isolated from ovine (strain km1h5-1p(t)) and bovine (strains ak-87, ocp and za-10(t)) rumen contents. based on 16s rrna gene sequence analysis, the methanogen strains were found to belong to the order methanobacteriales in the genus methanobrevibacter. strains za-10(t) and km1h5-1p(t) gained energy for growth by the reduction of co(2) to ch(4) using h(2) or formate exclusively as electron donors. increasing formate concentrations to 220 mm in batch culture ...200717329767
characterization of variation in rumen methanogenic communities under different dietary and host feed efficiency conditions, as determined by pcr-denaturing gradient gel electrophoresis analysis.understanding ruminal methanogens is essential for greenhouse gas mitigation, as well as for improving animal performance in the livestock industry. it has been speculated that ruminal methanogenic diversity affects host feed efficiency and results in differences in methane production. this study examined methanogenic profiles in the rumen using culture-independent pcr-denaturing gradient gel electrophoresis (pcr-dgge) analysis for 56 beef cattle which differed in feed efficiency, as well as die ...201020418436
methanogen community structure in the rumens of farmed sheep, cattle and red deer fed different diets.development of inhibitors and vaccines that mitigate rumen-derived methane by targeting methanogens relies on knowledge of the methanogens present. we investigated the composition of archaeal communities in the rumens of farmed sheep (ovis aries), cattle (bos taurus) and red deer (cervus elaphus) using denaturing gradient gel electrophoresis (dgge) to generate fingerprints of archaeal 16s rrna genes. the total archaeal communities were relatively constant across species and diets, and were less ...201121255054
a 'universal' type ii chaperonin pcr detection system for the investigation of archaea in complex microbial communities.bacteria and archaea are evolutionarily and biochemically distinct domains found together in many environments. robust 'universal' pcr primer sets targeting both the bacterial 16s rrna gene and the type i chaperonin gene have been established. however, 'universal' pcr primers for archaea are currently limited to the 16s rrna gene. we investigated the type ii chaperonin (known as the thermosome, tf55, cct or tcp-1) as a potential universal target (ut) for archaea. reproducible amplification of th ...201121776031
differences in the rumen methanogen populations of lactating jersey and holstein dairy cows under the same diet regimen.in the dairy cattle industry, holstein and jersey are the breeds most commonly used for production. they differ in performance by various traits, such as body size, milk production, and milk composition. with increased concerns about the impact of agriculture on climate change, potential differences in other traits, such as methane emission, also need to be characterized further. since methane is produced in the rumen by methanogenic archaea, we investigated whether the population structure of m ...201121705541
a1ao-atp synthase of methanobrevibacter ruminantium couples sodium ions for atp synthesis under physiological conditions.an unresolved question in the bioenergetics of methanogenic archaea is how the generation of proton-motive and sodium-motive forces during methane production is used to synthesize atp by the membrane-bound a(1)a(o)-atp synthase, with both proton- and sodium-coupled enzymes being reported in methanogens. to address this question, we investigated the biochemical characteristics of the a(1)a(o)-atp synthase (mbbra(1)a(o)) of methanobrevibacter ruminantium m1, a predominant methanogen in the rumen. ...201121953465
effect of dna extraction methods and sampling techniques on the apparent structure of cow and sheep rumen microbial communities.molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. therefore, sampling and dna extraction methods that result in adequate yields of microbial dna that also accurately represents the microbial community are crucial. fifteen different methods were used to extract dna from cow and sheep rumen samples. the dna yield and quality, and its suitability for downstream pcr amplifications varied consi ...201324040342
the effect of dietary chlorella vulgaris supplementation on micro-organism community, enzyme activities and fatty acid profile in the rumen liquid of goats.microalgae might be considered as an alternative source of fat and/or protein for ruminant's diets. however, changes in populations of ruminal micro-organisms associated with biohydrogenation process, methane and ammonia production in response to microalgae dietary supplementation have not been well characterized. thus, 16 cross-bred goats were divided into two groups. each goat of both groups was fed individually with alfalfa hay and concentrates separately. the concentrates of the control grou ...201727184296
high-throughput dna sequencing of the moose rumen from different geographical locations reveals a core ruminal methanogenic archaeal diversity and a differential ciliate protozoal diversity.moose rumen samples from vermont, alaska and norway were investigated for methanogenic archaeal and protozoal density using real-time pcr, and diversity using high-throughput sequencing of the 16s and 18s rrna genes. vermont moose showed the highest protozoal and methanogen densities. alaskan samples had the highest percentages of methanobrevibacter smithii, followed by the norwegian samples. one norwegian sample contained 43 % methanobrevibacter thaueri, whilst all other samples contained < 10  ...201528348818
a c subunit with four transmembrane helices and one ion (na+)-binding site in an archaeal atp synthase: implications for c ring function and structure.the ion-driven membrane rotors of atp synthases consist of multiple copies of subunit c, forming a closed ring. subunit c typically comprises two transmembrane helices, and the c ring features an ion-binding site in between each pair of adjacent subunits. here, we use experimental and computational methods to study the structure and specificity of an archaeal c subunit more akin to those of v-type atpases, namely that from pyrococcus furiosus. the c subunit was purified by chloroform/methanol ex ...201223007388
response of the rumen microbiota of sika deer (cervus nippon) fed different concentrations of tannin rich plants.high throughput sequencing was used to examine the rumen microbiota of sika deer fed high (olh) and low concentration (oll) of tannin rich oak leaves. the results showed that prevotella spp. were the most dominant bacteria. the most predominant methanogens were the members of the order methanoplasmatales. the dominant rumen protozoa were entodinium longinucleatum, eudiplodinium maggii, and epidinium caudatum, and the fungal communities were mostly represented by piromyces spp. moreover, the rela ...201525955033
effect of dietary fiber on the methanogen community in the hindgut of lantang gilts.the primary objective of this study was to investigate the effect of dietary fiber on methanogenic diversity and community composition in the hindgut of indigenous chinese lantang gilts to explain the unexpected findings reported earlier that lantang gilts fed low-fiber diet (lfd) produced more methane than those fed high-fiber diet (hfd). in total, 12 lantang gilts (58.7±0.37 kg) were randomly divided into two dietary groups (six replicates (pigs) per group) and fed either lfd (ndf=201.46 g/kg) ...201627052363
the complete genome sequence of the rumen methanogen methanobrevibacter millerae sm9.methanobrevibacter millerae sm9 was isolated from the rumen of a sheep maintained on a fresh forage diet, and its genome has been sequenced to provide information on the phylogenetic diversity of rumen methanogens with a view to developing technologies for methane mitigation. it is the first rumen isolate from the methanobrevibacter gottschalkii clade to have its genome sequence completed. the 2.54 mb sm9 chromosome has an average g + c content of 31.8 %, encodes 2269 protein-coding genes, and h ...201627536339
fiber degradation potential of natural co-cultures of neocallimastix frontalis and methanobrevibacter ruminantium isolated from yaks (bos grunniens) grazing on the qinghai tibetan plateau.several natural anaerobic fungus-methanogen co-cultures have been isolated from rumen and feces source of herbivores with strong fiber degrading ability. in this study, we isolated 7 neocallimastix with methanogen co-cultures from the rumen of yaks grazing on the qinghai tibetan plateau. based on morphological characteristics and internal transcribed spacer 1 sequences (its1), all the fungi were identified as neocallimastix frontalis. the co-cultures were confirmed as the one fungus - one methan ...201626979345
isolation, identification and fibrolytic characteristics of rumen fungi grown with indigenous methanogen from yaks (bos grunniens) grazing on the qinghai-tibetan plateau.to obtain co-cultures of anaerobic fungi and their indigenously associated methanogens from the rumen of yaks grazing on the qinghai-tibetan plateau and investigate their morphology features and ability to degrade lignocellulose.201626910857
vaccination of cattle with a methanogen protein produces specific antibodies in the saliva which are stable in the rumen.methane is produced in the rumen of cattle by a group of archaea (single-celled organisms forming a domain distinct from bacteria and eucarya) called methanogens. vaccination against methanogens has the potential to reduce methane emissions by inducing antibodies in saliva which are transferred to the rumen and diminish the ability of methanogens to produce methane. since it is likely that an effective vaccination strategy will need to produce high levels of methanogen-specific antibody in the s ...201525782351
methanogenic population and ch₄ production in swedish dairy cows fed different levels of forage.methanogenic community structure, methane production (ch(4)), and volatile fatty acid (vfa) profiles were investigated in swedish dairy cows fed a diet with a forage/concentrate ratio of 500/500 or 900/100 g/kg of dry matter (dm) of total dm intake (dmi). the rumen methanogenic population was evaluated using terminal restriction fragment length polymorphism (t-rflp) analysis, 16s rrna gene libraries, and quantitative real-time pcr (qrt-pcr). mean ch(4) yields did not differ (p > 0.05) between di ...201222752163
diversity, abundance and novel 16s rrna gene sequences of methanogens in rumen liquid, solid and epithelium fractions of jinnan cattle.three methanogen 16s rrna gene clone libraries were constructed from liquid (lm), solid (sm) and epithelium (em) fractions taken from the rumen of jinnan cattle in china. after the amplification by pcr using methanogen-specific primers met86f and met1340r, equal quantities of pcr products from the same fractions from each of the four cattle were mixed together and used to construct the three libraries. sequence analysis showed that the 268 lm clones were divided into 35 phylotypes with 18 sequen ...201022443614
intergenomic evolution and metabolic cross-talk between rumen and thermophilic autotrophic methanogenic archaea.methanobrevibacter ruminantium m1 (mru) is a rumen methanogenic archaean that can be able to utilize formate and co2/h2 as growth substrates. extensive analysis on the evolutionary genomic contexts considered herein to unravel its intergenomic relationship and metabolic adjustment acquired from the genomic content of methanothermobacter thermautotrophicus δh. we demonstrated its intergenomic distance, genome function, synteny homologs and gene families, origin of replication, and methanogenesis ...201727864137
technical note: comparison of biomarker and molecular biological methods for estimating methanogen abundance.quantitative real-time pcr (qpcr) has become a popular method for estimation of methanogen abundance in the ruminant digestive tract. however, there is no established method in terms of primer choice and quantification, which means that results are variable and not directly comparable between studies. archaeol has been proposed as an alternative marker for methanogen abundance, as it is ubiquitous in methanogenic archaea, and can be quantified by gas chromatography-mass spectrometry (gc-ms). the ...201324146154
plane of nutrition affects the phylogenetic diversity and relative abundance of transcriptionally active methanogens in the bovine rumen.methane generated during enteric fermentation in ruminant livestock species is a major contributor to global anthropogenic greenhouse gas emissions. a period of moderate feed restriction followed by ad libitum access to feed is widely applied in cattle management to exploit the animal's compensatory growth potential and reduce feed costs. in the present study, we utilised microbial rna from rumen digesta samples to assess the phylogenetic diversity of transcriptionally active methanogens from fe ...201729026096
characterization of natural co-cultures of piromyces with methanobrevibacter ruminantium from yaks grazing on the qinghai-tibetan plateau: a microbial consortium with high potential in plant biomass degradation.anaerobic fungi reside in the gut of herbivore and synergize with associated methanogenic archaea to decompose ingested plant biomass. despite their potential for use in bioconversion industry, only a few natural fungus-methanogen co-cultures have been isolated and characterized. in this study we identified three co-cultures of piromyces with methanobrevibacter ruminantium from the rumen of yaks grazing on the qinghai tibetan plateau. the representative co-culture, namely (piromyces + m. ruminan ...201728789484
taxon abundance, diversity, co-occurrence and network analysis of the ruminal microbiota in response to dietary changes in dairy cows.the ruminal microbiome, comprising large numbers of bacteria, ciliate protozoa, archaea and fungi, responds to diet and dietary additives in a complex way. the aim of this study was to investigate the benefits of increasing the depth of the community analysis in describing and explaining responses to dietary changes. quantitative pcr, ssu rrna amplicon based taxa composition, diversity and co-occurrence network analyses were applied to ruminal digesta samples obtained from four multiparous nordi ...201728704445
an adhesin from hydrogen-utilizing rumen methanogen methanobrevibacter ruminantium m1 binds a broad range of hydrogen-producing microorganisms.symbiotic associations are ubiquitous in the microbial world and have a major role in shaping the evolution of both partners. one of the most interesting mutualistic relationships exists between protozoa and methanogenic archaea in the fermentative forestomach (rumen) of ruminant animals. methanogens reside within and on the surface of protozoa as symbionts, and interspecies hydrogen transfer is speculated to be the main driver for physical associations observed between the two groups. in silico ...201626643468
annotating functional rnas in genomes using infernal.many different types of functional non-coding rnas participate in a wide range of important cellular functions but the large majority of these rnas are not routinely annotated in published genomes. several programs have been developed for identifying rnas, including specific tools tailored to a particular rna family as well as more general ones designed to work for any family. many of these tools utilize covariance models (cms), statistical models of the conserved sequence, and structure of an r ...201424639160
the crystal structure of methenyltetrahydromethanopterin cyclohydrolase from methanobrevibacter ruminantium.methenyltetrahydromethanopterin cyclohydrolase (mch) is involved in the methanogenesis pathway of archaea as a c1 unit carrier where n(5) -formyl-tetrahydromethanopterin is converted to methenyl-tetrahydromethanopterin. mch from methanobrevibacter ruminantium was cloned, purified, crystallized and its crystal structure solved at 1.37 å resolution. a biologically active trimer, the enzyme is composed of two domains including an n-terminal domain of six α-helices encompassing a series of four β-sh ...201323873651
the effect of saturated fatty acids on methanogenesis and cell viability of methanobrevibacter ruminantium.saturated fatty acids (sfas) are known to suppress ruminal methanogenesis, but the underlying mechanisms are not well known. in the present study, inhibition of methane formation, cell membrane permeability (potassium efflux), and survival rate (live/dead staining) of pure ruminal methanobrevibacter ruminantium (dsm 1093) cell suspensions were tested for a number of sfas. methane production rate was not influenced by low concentrations of lauric (c12; 1 μg/ml), myristic (c14; 1 and 5 μg/ml), or ...201323710130
methane production in dairy cows correlates with rumen methanogenic and bacterial community structure.methane (ch4) is produced as an end product from feed fermentation in the rumen. yield of ch4 varies between individuals despite identical feeding conditions. to get a better understanding of factors behind the individual variation, 73 dairy cows given the same feed but differing in ch4 emissions were investigated with focus on fiber digestion, fermentation end products and bacterial and archaeal composition. in total 21 cows (12 holstein, 9 swedish red) identified as persistent low, medium or h ...201728261182
few highly abundant operational taxonomic units dominate within rumen methanogenic archaeal species in new zealand sheep and cattle.sequencing and analyses of 16s rrna gene amplicons were performed to estimate the composition of the rumen methanogen community in 252 samples from eight cohorts of sheep and cattle, separated into 16 different sample groups by diet, and to determine which methanogens are most prominent in the rumens of farmed new zealand ruminants. methanobacteriales (relative abundance ± standard deviation, 89.6% ± 9.8%) and methanomassiliicoccales (10.4% ± 9.8%) were the two major orders and contributed 99.98 ...201525416771
a high-throughput screening assay for identification of inhibitors of the a1ao-atp synthase of the rumen methanogen methanobrevibacter ruminantium m1.we report the development of a high-throughput screening platform to identify inhibitors of the membrane-bound a1ao-atp synthase from the rumen methanogen methanobrevibacter ruminantium m1. inhibitors identified in the screen were tested against growing cultures of m. ruminantium, validating the approach to identify new inhibitors of methanogens.201525575416
influence of ph and the degree of protonation on the inhibitory effect of fatty acids in the ruminal methanogen methanobrevibacter ruminantium strain m1.to investigate the relationship between the protonation of medium-chain fatty acids (mcfa) and their inhibitory effect on a ruminal methanogen species.201526394014
phloroglucinol degradation in the rumen promotes the capture of excess hydrogen generated from methanogenesis inhibition.strategies to manage metabolic hydrogen ([h]) in the rumen should be considered when reducing ruminant methane (ch4) emissions. however, little is known about the use of dietary treatments to stimulate rumen microorganisms capable of capturing the [h] available when ch4 is inhibited in vivo. the effects of the phenolic compound phloroglucinol on ch4 production, [h] flows and subsequent responses in rumen fermentation and microbial community composition when methanogenesis is inhibited were inves ...201729051749
inhibition of methyl-com reductase from methanobrevibacter ruminantium by 2-bromoethanesulfonate.cattle husbandry is a major contributor to atmospheric methane, which is considered as an important greenhouse gas. moreover, the generation of methane in the intestine of domestic ruminants by methanogenic bacteria is a drag on feed efficacy. studies on methanogenesis have typically implied model organisms that are, however, not relevant in the ruminant gut. this paper shows that methyl-com reductase catalyzing the final step of methanogenesis in methanobrevibacter ruminantium, a major particip ...201425483006
methanogenic archaea diversity in hanwoo (bos taurus coreanae) rumen fluid, rectal dung, and barn floor manure using a culture-independent method based on mcra gene sequences.the diversity of methanogenic archaea associated with korean hanwoo cattle was analyzed using mcra gene sequences from samples of rumen fluid (rf), rectal dung (rd), and barn floor manure (bfm). the predominant species were methanobrevibacter ruminantium in rf and bfm(63.6% and 62.4%, respectively) and methanocorpusculum labreanum in rd (53.2%).201424508643
metatranscriptomic profiling reveals linkages between the active rumen microbiome and feed efficiency in beef cattle.exploring compositional and functional characteristics of the rumen microbiome can improve the understanding of its role in rumen function and feed efficiency. in this study, we applied metatranscriptomics to characterize the active rumen microbiomes of beef cattle with different feed efficiencies (efficient, n=10; inefficient, n=10) using total rna sequencing. active bacterial and archaeal compositions were estimated based on 16s rrnas, and active microbial metabolic functions including carbohy ...201728235871
differences in the methanogen population exist in sika deer (cervus nippon) fed different diets in china.understanding the methanogen structure from sika deer (cervus nippon) in china may be beneficial to methane mitigation. in the present preliminary study, we investigated the methanogen community in the rumen of domesticated sika deer fed either tannin-rich plants (oak leaf, ol group) or corn stalk (cs group) using 16s rrna gene clone libraries. overall, we obtained 197 clone sequences, revealing 146 unique phylotypes, which were assigned to 36 operational taxonomic units at the species level (98 ...201324061342
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